Method of making an artificial micro-gland using taxis

Abstract

A method is used for making an artificial micro-gland by taxis. A monodisperse multiple emulsion is produced with a first fluid; a second fluid confined within the first fluid; a third fluid within the second fluid. Interfaces between the fluids permit living cells dispersed in the one of the fluids to migrate towards an adjacent fluid having a different concentration of an agent affecting the metabolic activity of the living cells. Waiting, usually about 30 minutes, allows the living cells to migrate to the interface, forming the continuous membrane. Once formed, the artificial micro-gland is removed from the remains of the emulsion. The artificial micro-gland may also be given a second layer of different cells when the emission of the cells of the artificial micro-gland is used as the agent to attract the different cells. The method may also be used to produce an artificial micro-gland within an artificial micro-gland.

What is claimed is: 
     
         1 . A method of making an artificial micro-gland by taxis, the artificial micro-gland comprising a continuous membrane of living cells, the continuous membrane defining an enclosed volume, the enclosed volume comprising a reservoir serving as a bioreactor, the method comprising the steps of:
 producing a monodisperse multiple emulsion, the monodisperse multiple emulsion comprising:
 a first fluid serving as a host environment; 
 a second fluid confined within the host environment, the second fluid being immiscible in the first fluid; 
 a third fluid within the second fluid, the third fluid being immiscible in the second fluid such that there is an interface between the second fluid and the third fluid, the third fluid comprising a plurality of living cells dispersed therein, said living cells capable of metabolic activity; and, 
 an agent capable of affecting the metabolic activity of the living cells, the agent present within the second fluid at a higher concentration than in the third fluid; 
   waiting until the living cells migrate to the interface between the second fluid and the third fluid to form the continuous membrane around the third fluid; and,   removing the first fluid and the second fluid from the monodisperse multiple emulsion to produce the artificial micro-gland.   
     
     
         2 . The method of  claim 1 , wherein the:
 first fluid of the host environment comprises water;   second fluid comprises oil;   third fluid comprises water;   agent is selected from the group consisting of: oxygen; carbon dioxide; nitrogen oxide; sugar; phosphates, nitrates, sulphates, and potassium salts; cyclic adenosine monophosphate (CAMP); inositon phospholipid (mPIP3); actin; histamine; serotonin (5HT); plaletet acting factors (PAF); arachidonic acid metabolites; diacykglyseril (IP3); leukotine B4; lipoxins; prostaglandins; cytotaxin; f-met-leu-phe tripeptide; cytokines; kinins,cytotaxins; anaphylatoxin peptide (C5a); aspartic acid (ASP); serine (SER); and, chemo-attractants; and,   living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells.   
     
     
         3 . A method of making an artificial micro-gland by taxis, the artificial micro-gland comprising a continuous membrane of living cells, the continuous membrane defining an enclosed volume, the enclosed volume comprising a reservoir serving as a bioreactor, the method comprising the steps of:
 producing a monodisperse multiple emulsion, the monodisperse multiple emulsion comprising:
 a first fluid serving as a host environment; 
 a second fluid within the host environment, the second fluid comprising a plurality of living cells dispersed therein, said living cells capable of metabolic activity; 
 a third fluid within the second fluid being immiscible in the second fluid such that there is an interface between the second fluid and the third fluid; 
 an agent capable of affecting the metabolic activity of the living cells, the agent present within the third fluid at a higher concentration than in the second fluid; 
 wherein the first fluid serving as the host environment maintains a relatively low concentration of the agent compared to the third fluid; and, 
   waiting until the living cells to migrate to the interface between the second fluid and the third fluid to form the continuous membrane around the third fluid; and,   removing the first fluid and the second fluid from the multiple emulsion to produce the artificial micro-gland,   
     
     
         4 . The method of  claim 3 , wherein the:
 first fluid comprises a first oil;   second fluid comprises water;   third fluid comprises a second oil;   agent is selected from the group consisting of: oxygen; carbon dioxide; nitrogen oxide; sugar; phosphates, nitrates, sulphates, and potassium salts; cyclic adenosine monophosphate (cAMP); inositon phospholipid (mPIP3); actin; histamine; serotonin (5HT); plaletet acting factors (PAF); arachidonic acid metabolites; diacykglyseril (IP3); leukotine B4; lipoxins; prostaglandins; cytotaxia; f-met-leu-phe tripeptide; cytokines; kinins,cytotaxins; anaphylatoxin peptide (C5a); aspartic acid (ASP); serine (SER); and chemo-attractants; and,   living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells.   
     
     
         5 . A method of making an artificial micro-gland by taxis, the artificial micro-gland comprising a continuous membrane of living cells, the continuous membrane defining an enclosed volume, the enclosed volume comprising a reservoir serving as a bioreactor, the method comprising the steps of:
 producing a monodisperse multiple emulsion, the monodisperse multiple emulsion comprising:
 a first fluid serving as a host environment; 
 a second fluid within the host environment, the second fluid being immiscible in the first fluid such that there is an interface between the first fluid and the second fluid, the second fluid comprising a plurality of living cells dispersed therein, said living cells capable of metabolic activity; 
 a third fluid within the second fluid; and, 
 an agent capable of affecting the metabolic activity of the living cells, the agent present within the first fluid at a higher concentration than in the second fluid and in the third fluid; 
   waiting until the living cells migrate to the interface between the first fluid and the second fluid to form the continuous membrane around the second fluid; and,   removing the first fluid from the monodisperse multiple emulsion to produce the artificial micro-gland.   
     
     
         6 . The method of  claim 5 , wherein the:
 first fluid comprises a first oil;   second fluid comprises water;   third fluid comprises a second oil;   agent is selected from the group consisting of: oxygen; carbon dioxide; nitrogen oxide; sugar; phosphates, nitrates, sulphates, and potassium salts; cyclic adenosine monophosphate (cAMP); inositon phospholipid (mPIP3); actin; histamine; serotonin (5HT); plaletet acting factors (PAF); arachidonic acid metabolites; diacykglyseril (IP3); leukotine B4; lipoxins; prostaglandins; cytotaxin; f-met-leu-phe tripeptide; cytokines; kinins,cytotaxins; anaphylatoxin peptide (C5a); aspartic acid (ASP); serine (SER); and, chemo-attractants; and,   the living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells.   
     
     
         7 . A method of making a multi-shell artificial micro-gland by taxis, the multi-shell artificial micro-gland comprising a plurality of continuous membranes of first living cells, the continuous membranes defining an enclosed volume, the enclosed volume comprising a reservoir serving as a bioreactor, the method comprising the steps of:
 producing a monodisperse emulsion, the monodisperse emulsion comprising:
 a fluid serving as a host environment; 
 an artificial micro-gland within the host environment, the artificial micro-gland comprising a first continuous membrane of first living cells surrounding a reservoir; and, 
 a plurality of second living cells dispersed within the fluid, said second living cells capable of metabolic activity stimulated by discharges of an agent from the first living cells; 
   waiting until the second living cells migrate to the living cells to form a second continuous membrane covering the first living cells; and,   removing the fluid from the emulsion to produce the multi-shell artificial micro-gland.   
     
     
         8 . The method of  claim 7 , wherein the:
 fluid comprises oil or water;   first living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells; and,   second living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells, provided that the selected second living cells are different than the first living cells.   
     
     
         9 . A method of making an artificial micro-gland by taxis, the artificial micro-gland comprising a first artificial micro-gland within a second artificial micro-gland, the method comprising the steps of:
 producing a monodisperse multiple emulsion, the monodisperse multiple emulsion comprising:
 a first fluid serving as a host environment; 
 a second fluid confined within the host environment, the second fluid being immiscible in the first fluid such that there is a first interface between the first fluid and the second fluid; 
 a third fluid within the second fluid, the third fluid being immiscible in the second fluid such that there is a second interface between the second fluid and the third fluid, the third fluid comprising a plurality of first living cells dispersed therein, said first living cells capable of metabolic activity; and, 
 an agent capable of affecting the metabolic activity of the living cells, the agent present within the second fluid at a higher concentration than in the third fluid; 
   waiting until the first living cells migrate to the second interface between the second fluid and the third fluid to form a continuous membrane around the third fluid, forming a first artificial micro-gland within the second fluid;   adding second living cells to the first fluid;   waiting until the second living cells migrate to the first interface between the first fluid and the second fluid to form a second continuous membrane covering the second fluid and forming the second artificial micro-gland; and,   removing the first fluid from the monodisperse multiple emulsion to produce the artificial micro-gland comprising a first artificial micro-gland within a second artificial micro-gland.   
     
     
         10 . The method of  claim 9 , wherein the:
 first fluid comprises water;   second fluid comprises oil;   third fluid comprises water;   agent is selected from the group consisting of: oxygen; and, carbon dioxide;   first living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells; and;   second living cells are selected from the group consisting of: eukaryotic cells; and; prokaryotic cells.